EP3534932B1 - Traitement de maladies associées à l'igfb3 et à son récepteur - Google Patents

Traitement de maladies associées à l'igfb3 et à son récepteur Download PDF

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EP3534932B1
EP3534932B1 EP17866859.6A EP17866859A EP3534932B1 EP 3534932 B1 EP3534932 B1 EP 3534932B1 EP 17866859 A EP17866859 A EP 17866859A EP 3534932 B1 EP3534932 B1 EP 3534932B1
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tmem219
cancer
igfbp
agonist
seq
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EP3534932A1 (fr
EP3534932A4 (fr
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Youngman Oh
Mikhail Dozmorov
Melissa Qing CAI
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Virginia Commonwealth University
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Virginia Commonwealth University
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • G01N33/57492—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds localized on the membrane of tumor or cancer cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001102—Receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/75—Agonist effect on antigen
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/71—Assays involving receptors, cell surface antigens or cell surface determinants for growth factors; for growth regulators
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

  • the invention generally relates to methods of treating diseases involving insulin-like growth factor-binding protein 3 (IGFBP-3) and its receptor, IGFBP-3R using IGFBP-3R agonists.
  • IGFBP-3 insulin-like growth factor-binding protein 3
  • TNBC triple negative breast cancer
  • Metabolic syndrome is a serious health condition that is becoming more and more prevalent as frequency of obesity and sedentary lifestyles arise, and as a result of aging populations. For example, in the United States, about 34% of the population has metabolic syndrome, and the prevalence increases with age: metabolic syndrome affects about 60% of the U.S. population older than age 50. Metabolic syndrome is associated with an increased risk of several debilitating diseases, including insulin resistance, atherosclerotic cardiovascular disease (e.g., heart disease and stroke) and type 2 diabetes. The development of these diseases results in a high negative impact on the quality of life of those who are afflicted, and places a high burden on the already strained health care systems of countries. While some treatments are available for specific symptoms (e.g. drugs for high blood pressure, etc.), and while life style changes can have a positive impact, all patients do not respond equally well to medications or to the need for life style changes. It would be beneficial to have available additional medicaments to treat metabolic syndrome.
  • specific symptoms e.g. drugs for high blood pressure,
  • Obstructive respiratory disorders also known as obstructive lung or pulmonary disease
  • COPD chronic obstructive pulmonary disease
  • asthma a category of respiratory disease characterized by airway obstruction.
  • COPD chronic obstructive pulmonary disease
  • the incidence of these maladies is on the rise.
  • COPD chronic obstructive pulmonary disease
  • the disease burden and its financial impact is predicted to increase, e.g. due to population aging.
  • As of 2014 it was estimated that asthma affected as many as 334 million people worldwide. It is the most common chronic disease in children and its prevalence is also rising. While there are some medications available to control symptoms, there is an ongoing need to provide additional improved methods and agents for treating these and other types of obstructive respiratory disorders.
  • WO201216158 discloses a link between low levels of IGFBP-3R (TMEM-219) and certain diseases, and provides agonistic polyclonal antibodies against this factor.
  • IGFBP-3R critical antitumor, anti-inflammatory signaling cascade, the IGFBP-3/IGFBP-3R axis. It has been discovered that IGFBP-3 and its receptor, IGFBP-3R play a role in several diseases, including cancer, metabolic syndrome, obstructive respiratory disorders and various inflammatory disorders. (IGFBP-3R is also known as "transmembrane protein 219" and “IGFBP-3R” and the acronym “TMEM219” may be used interchangeably herein.) For such diseases, in some aspects, it has been determined that the level of IGFBP-3 that is produced is insufficient to cause sufficient activation of IGFBP-3R (TMEM219). Therefore, the present disclosure provides agents that substitute for the natural ligand IGFBP-3.
  • the agents are TMEM219 agonists which bind to and activate TMEM219 and can thus be used to prevent, treat or ameliorate symptoms of such diseases and/or in some cases, the recurrence of the diseases, and/or improve the prognosis (e.g. survival rate, rate of relapse, disease free survival time, etc.) of patients suffering from the diseases.
  • the TMEM219 agonists are monoclonal antibodies (mAbs).
  • mAbs monoclonal antibodies
  • TMEM219 agonistic antibodies constitute a new generation of therapeutics with a unique mechanism and target specificity for treating these disorders.
  • the TMEM219 agonist mAbs advantageously exhibit no deleterious harmful effects (such as cell damage or cell killing) on normal, non-disease (e.g. non-tumor) cells.
  • TMEM219 in tumor cells is indicative of a poor prognosis, e.g. an increased risk of metastasis, recurrence and/or a lower overall chance of survival.
  • an aggressive treatment regimen is typically recommended for patients with levels of expression of TMEM219 that are lower than a predetermined, corresponding reference value.
  • patients with a high level of expression of TMEM219 in tumor cells have a relatively good prognosis, with a lower risk of metastasis and recurrence and/or a higher chance of survival. Accordingly, a less aggressive (and thus less toxic) treatment regimen with fewer side effects is recommended.
  • the agonist is an antibody that binds to and activates IGFBP-3R and comprises:
  • the antibody comprises i) a heavy chain variable region with an amino acid sequence that is at least 90% identical to SEQ ID NO: 6, and ii) a light chain variable region with an amino acid sequence that is at least 90% identical to SEQ ID NO: 8.
  • the antibody comprises wherein the heavy chain variable region of said antibody comprises the sequence as set forth in SEQ ID NO: 6 and wherein the light chain variable region of said antibody comprises the sequence as set forth in SEQ ID NO: 8.
  • the antibody comprises a detectable label.
  • An aspect disclosed but not covered by the subject matter of the claims comprises a method of determining a prognosis of a subject with cancer and treating the subject accordingly, comprising, i) measuring a level of TMEM219 expression in a tumor sample from the subject ii) comparing the level of TMEM219 expression obtained in step i) with a corresponding reference level of TMEM219 expression; and iii) if the level of TMEM219 expression is the same or lower than the corresponding reference level of TMEM219 expression, then iv) concluding that the patient has a poor prognosis and providing an aggressive anti-cancer treatment to the patient; or v) if the level of TMEM219 expression is higher than the corresponding reference level of TMEM219 expression, then iv) concluding that the patient has a good prognosis and providing a less aggressive anti-cancer treatment to the patient.
  • the cancer is breast cancer, colon cancer, lung cancer, ovarian cancer, pancreatic cancer, liver cancer or leukemia.
  • the prognosis includes one or more of risk of recurrence of the cancer in the patient, risk of metastasis, overall survival of the patient and prediction of the benefit of chemotherapy for the patient.
  • the present disclosure describes therapeutic agents and methods of their use to treat diseases involving IGFBP-3 and its receptor, IGFBP-3R, e.g. diseases and conditions caused by abnormal functioning of IGFBP-3 and IGFBP-3R.
  • IGFBP-3R diseases and conditions caused by abnormal functioning of IGFBP-3 and IGFBP-3R.
  • agents that bind to and activate IGFBP-3R i.e. IGRBP-3R agonists
  • IGFBP-3 substitutes agents that bind to and activate IGFBP-3R (i.e. IGRBP-3R agonists) are used as IGFBP-3 substitutes.
  • IGFBP-3 substitutes agents that bind to and activate IGFBP-3R (i.e. IGRBP-3R agonists) are used as IGFBP-3 substitutes.
  • IGFBP-3 substitutes agents that bind to and activate IGFBP-3R (i.e. IGRBP-3R agonists) are used as IGFBP-3 substitutes.
  • IGFBP-3 substitutes agents that bind to and activate
  • the level of expression of IGFBP-3R in tumor cells is used as an indicator of the prognosis of a cancer patient, with low levels indicating a poor prognosis and high levels indicating a relatively good prognosis. This type of assessment allows medical practitioners to tailor recommended cancer treatment regimens on a patient by patient basis.
  • IGFBP-3R or "TMEM219” refers to the Homo sapiens (human) protein that acts as the receptor for human "insulin-like growth factor-binding protein 3" or "IGFBP-3".
  • the receptor is also known as “transmembrane protein 219” and is encoded by the TMEM219 gene (gene ID 124446 in the NCBI database).
  • agonist we mean a chemical (compound, substance, etc.) that binds to a receptor and activates the receptor to produce a biological response.
  • the agonists are monoclonal antibodies (mAbs) specific for binding to the receptor "IGFBP-3R". Upon binding, the mAbs activate the receptor, i.e. its biological activity is elicited, increased, etc., compared to the level of activation when no agonist or natural ligand is bound.
  • a “therapeutically effective amount” of a compound is an amount that is sufficient to treat or prevent or ameliorate (lessen) at least one symptom of a disease.
  • treat or “treating” a disease, we mean that, in a treated individual, one or more unwanted symptoms of the disease is/are eliminated (i.e. the patient is cured), or lessened, and/or the time interval during which the symptoms are present is shortened, and/or onset of symptoms is delayed, compared to an untreated individual.
  • Prevention refers to stopping or averting (warding off, etc.) the occurrence of a disease or a disease aspect or symptom before it occurs, e.g. before evidence of the disease, symptom, etc. is detectable or measurable.
  • VH CDR refers to a heavy chain variable domain complementarity determining region (CDR) of an antibody.
  • LH CDR refers to a light chain variable domain CDR of an antibody.
  • CDR1, CDR2 and CDR3 There are three CDRs (CDR1, CDR2 and CDR3), arranged non-consecutively, on the amino acid sequence of a variable domain of an antigen receptor. Since the antigen receptors are typically composed of two variable domains (on two different polypeptide chains, one heavy chain and one light chain), there are six CDRs for each antigen receptor that can collectively come into contact with the antigen.
  • a single antibody molecule has two antigen receptors and therefore contains a total of twelve CDRs, although sixty CDRs are found on a pentameric IgM molecule.
  • the agonists of IGFBP-3R of the invention are characterized by the heavy chain and light chain CDRs as defined in the claims.
  • agonists capable of activating the IGFBP-3R defined in more general terms. They may be molecules that bind to the receptor, for example at the IGFBP-3 binding site which is described below, specifically or selectively.
  • the agonists may be of any of the many known types of molecules which bind to receptors, for example small molecule drugs, antibodies, etc.
  • mAbs monoclonal antibodies
  • the mAbs may be used to deliver the CDRs to the receptor (i.e.
  • antibodies may be used to mediate contact between one or more CDRs and the receptor binding site) other molecules which contain one or more of the CDRs may also be used to do so, e.g. peptides and polypeptides which comprise one or more CDRs.
  • peptides and polypeptides may be protected to decrease proteolysis and increase bioavailability, e.g. by including "non-natural" or non-cleavable amino acids (e.g.
  • the agonist may be a small molecule drug which fits the receptor binding site and binds sufficiently to activate the receptor.
  • small molecule drug we mean an organic compound that is of a low molecular weight ( ⁇ 900 daltons) and which has a size on the order of 1 nm.
  • agonists typically bind to target receptors via one or more of electrostatic bonding, hydrogen bonding, and/or van der Waals/London dispersion forces.
  • the agonists that are not antibodies bind to or within residues 116-125 of IGFBP-3R, the amino acid sequence of which is GLKGSSAGQL (SEQ ID NO: 13), as described below for the monoclonal antibodies
  • agonists may be monoclonal antibodies (mAbs) specific for binding to and activating IGFBP-3R.
  • ED 50 effective dose, causing 50% of maximum effect for the measured biological effects in cells receiving the drug
  • the mAbs generally exhibit an ED50 in the range of from about 1 - 100 nM, e.g. about 5 to 50 nM such as about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nM or more.
  • the mAbs may or may not bind to the exact same residues that are bound by the natural ligand; however, they bind sufficiently to activate the receptor. In some aspects, the antibodies bind to the same site at which the ligand IGFBP-3 binds. Whatever the exact position of binding, the mAbs stand in for/make up for the lack of natural ligand binding and once bound, they activate the receptor. In some aspects, the mAbs bind, for example, to portions of IGFBP-3R which are accessible and not buried in the membrane, e.g. within residues 1-197 of IGFBP-3R.
  • the mAbs bind to or within residues 116-125 of IGFBP-3R, the amino acid sequence of which is GLKGSSAGQL (SEQ ID NO: 13). In other words, in some aspects, the mAbs bind to at least 1, 2, 3, 4, 5, 6, 7, 8, or 9 consecutive amino acids within SEQ ID NO: 13, or to all 10 amino acids of SEQ ID NO:13. In yet other aspects, the mAbs bind to from 1-9 amino acids which are not consecutive in sequence, i.e. one or more (e.g. about 1-9) amino acids within this sequence do not bind directly to the mAb. Binding to the mAbs is generally non-covalent, e.g. via one or more of electrostatic bonding, hydrogen bonding, and/or van der Waals/London dispersion forces.
  • Exemplary mAbs are described herein and the sequences of exemplary mAbs are shown in Figure 5 .
  • conservative and/or non-conservative amino acid substitutions may be made in the sequences as long as the resulting mAbs retain the ability to bind to and act as agonists of IGFBP-3R.
  • Such variants generally have at least about 50% identity to the sequences disclosed herein, e.g. at least about 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identity to the disclosed sequences.
  • antibodies may be designed and produced which contain one or more complementarity-determining regions (CDRs) of the antibodies described herein, i.e. they contain at least one paratope or antigen-binding region as described herein (such as at least one CDR), but contain different non-CDR sequences.
  • CDRs complementarity-determining regions
  • Other variations include but are not limited to: human, humanized, or chimeric antibodies, antibody fragments that bind IGFBP-3R (e.g. human IGFBP-3R), a Fab', a F(ab')2, a F(ab')3, a monovalent scFv, a bivalent scFv, a single domain antibody, etc.
  • the antibodies may be IgG, IgM, or IgA antibodies or antigen binding fragments thereof.
  • the antibodies may be labeled with a detectable label as described in detail below, or may be otherwise modified, e.g. by glycosylation. All such variants can be contemplated, so long as they bind to IGFBP-3R and acts as agonists of IGFBP-3R.
  • TMEM219 #245 TMEM219 #274 or TMEM219 #274-hIgG1 chimera.
  • the latter is a human IgG1 chimera of TMEM219 #274, which is the specific object of the invention.
  • Tables 3-5 show the CDR sequences of heavy and light chains of TMEM219 #245,-TMEM219 #274 and TMEM219 #274-hIgG1 chimera.
  • TMEM219#245 was compared to the germline IGHV9 gene sequence for the heavy chain CDRs and to the IGKV9 gene sequence for the light chain CDRs whereas TMEM219#274 heavy chain CDRs and light chain CDRs were compared to the IGHV9 and IGKV6 gene sequences, respectively.
  • CDR sequences of TMEM219 #274-hIgG1 chimera are the same as those of TMEM219 #274.
  • the CDRs may be denominated a first, second, third, etc. antibody as required for clarity. Table 1.
  • TMEM219 #274-hIgG1 chimera CDR sequences (same as those of TMEM219@274 _Heavy chain 6
  • Light chain CDR1-IMGT Protein G Y T F T N Y G (SEQ ID NO: 25) Protein: Q N V G T N (SEQ ID NO: 31)
  • CDR2-IMGT Protein IN T Y T R E T (SEQ ID NO: 27)
  • the antibody comprises one or more of the CDRs of antibody TMEM219 mAb#1 (#245) as follows: (a) a VH CDR at least 90% identical to SEQ ID NO: 15; (b) a VH CDR at least 90% identical to SEQ ID NO: 17; (c) a VH CDR at least 90% identical to SEQ ID NO: 19; (d) a VL CDR at least 90% identical to VL CDR1 of SEQ ID NO: 21; (e) a VL CDR at least 90% identical to SEQ ID NO: 23; and (f) a VL CDR at least 90% identical to the sequence ATS. That is, the sequence is about 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or even 100% identical to the indicated sequence.
  • the antibody comprises the CDRs of antibody TMEM219 mAb#2 (#274) as follows: (a) a heavy chain complementary determining region 1 having the sequence GYTFTFNYG (SEQ NO: 25), (b) a heavy chain complementary determining region 2 having the sequence INTYTRET (SEQ NO: 27), (c) a heavy chain complementary determining region 3 having the sequence ARGSTMYGLDK (SEQ NO: 29), (d) a light chain complementary determining region 1 having the sequence QNVGTN (SEQ NO: 31),(e) a light chain complementary determining region 2 having the sequence SAS, (f) a light chain complementary determining region 3 having the sequence HQYNSYPLT (SEQ NO: 33).
  • the heavy chain variable region of said antibody may comprise the sequence at least 90% identical to SEQ ID NO: 6 and the light chain variable region of said antibody may comprise the sequence at least 90% identical to SEQ ID NO: 8.
  • the heavy chain variable region of said antibody may comprise the sequence as set forth in SEQ ID NO: 6 and the light chain variable region of said antibody may comprise the sequence as set forth in SEQ ID NO: 8.
  • antibodies for use in the methods described herein are obtainable from hybridomas using technology that is known in the art.
  • the antibodies may be made by recombinant technology, e.g. by cell culture or bacterial culture, as is well-known in the art, or even synthetically via chemical peptide synthesis.
  • the antibodies disclosed can be used for treating diseases, specifically, the antibodies of the invention are for use in the treatment of cancer expressing IGFBP-3R.
  • the antibodies may be modified to include other effector molecules.
  • effector molecules that can be attached to antibodies include toxins, therapeutic enzymes, antibiotics, radio-labeled nucleotides and the like.
  • linking molecules may be used to join the antibody to the effector molecule. Such effectors may be especially useful when the disease that is treated is cancer.
  • nucleic acids encoding the aforementioned molecules are also disclosed; they need not be identical to those depicted in Figure 5A-F, e.g. due to the redundancy of the genetic code.
  • encoding sequences will generate the antibodies described herein, and may or may not be codon optimized for production in a particular way, e.g. in plant, mammalian or bacterial host cells.
  • Nucleic acids include but are not limited to DNA and RNA, and sequences that are at least about 90% homologous to the sequences disclosed herein (e.g. about 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or even 100% homologous).
  • vectors plasmids, cosmids, viral vectors, etc.
  • cells which contain the nucleic acid sequences and/or the vectors.
  • compositions comprising one or more of the agonists (e.g. antibodies) described herein.
  • the components in the compositions will vary depending on whether the antibodies are used in diagnostic methods or in treatment methods, whether or not they are labeled, etc.
  • compositions When used for treatment methods, the compounds described herein are generally delivered (administered) as a pharmaceutical composition/formulation.
  • the compositions generally include one or more substantially purified antibodies as described herein, and a pharmacologically suitable (physiologically compatible) carrier, which may be aqueous or oil-based.
  • a pharmacologically suitable (physiologically compatible) carrier which may be aqueous or oil-based.
  • such compositions are prepared as liquid solutions or suspensions, or as solid forms such as tablets, pills, powders and the like.
  • Solid forms suitable for solution in, or suspension in, liquids prior to administration are also contemplated (e.g. lyophilized forms of the compounds), as are emulsified preparations.
  • the liquid formulations are aqueous or oil-based suspensions or solutions.
  • the active ingredients are mixed with excipients which are pharmaceutically acceptable and compatible with the active ingredients, e.g. pharmaceutically acceptable salts.
  • Suitable excipients include, for example, water, saline, dextrose, glycerol, ethanol and the like, or combinations thereof.
  • the composition may contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents, preservatives, and the like. If it is desired to administer an oral form of the composition, various thickeners, flavorings, diluents, emulsifiers, dispersing aids or binders and the like are added.
  • the composition may contain any such additional ingredients so as to provide the composition in a form suitable for administration.
  • the final amount of antibody in the formulations varies, but is generally from about 1-99%. Still other suitable formulations are found, for example in Remington's Pharmaceutical Sciences, 22nd ed. (2012; eds. Allen, Adejarem Desselle and Felton).
  • materials which can serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as twin 80, phosphates, glycine, sorbic acid, or potassium sorbate), partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, or zinc salts), colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, methylcellulose, hydroxypropyl methylcellulose, wool fat, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
  • “Pharmaceutically acceptable salts” refers to the relatively non-toxic, inorganic and organic acid addition salts, and base addition salts, of compounds disclosed herein. These salts can be prepared in situ during the final isolation and purification of the compounds. In particular, acid addition salts can be prepared by separately reacting the purified compound in its free base form with a suitable organic or inorganic acid and isolating the salt thus formed.
  • Exemplary acid addition salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactiobionate, sulfamates, malonates, salicylates, propionates, methylene-bis-.beta.-hydroxynaphthoates, gentisates, isethionates, di-p-toluoyltartrates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, cyclohexylsulfamates and
  • Base addition salts can also be prepared by separately reacting the purified compound in its acid form with a suitable organic or inorganic base and isolating the salt thus formed.
  • Base addition salts include pharmaceutically acceptable metal and amine salts. Suitable metal salts include the sodium, potassium, calcium, barium, zinc, magnesium, and aluminum salts. The sodium and potassium salts are preferred.
  • Suitable inorganic base addition salts are prepared from metal bases which include sodium hydride, sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide, zinc hydroxide and the like.
  • Suitable amine base addition salts are prepared from amines which have sufficient basicity to form a stable salt, and preferably include those amines which are frequently used in medicinal chemistry because of their low toxicity and acceptability for medical use.
  • ammonia ethylenediamine, N-methyl-glucamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, diethylamine, piperazine, tris(hydroxymethyl)-aminomethane, tetramethylammonium hydroxide, triethylamine, dibenzylamine, ephenamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, ethylamine, basic amino acids, e.g., lysine and arginine, and dicyclohexylamine, and the like.
  • compositions may be administered in vivo by any suitable route including but not limited to: inoculation or injection (e.g. intravenous, intraperitoneal, intramuscular, subcutaneous, intra-aural, intraarticular, intramammary, intratumoral, and the like), by topical application and by absorption through epithelial or mucocutaneous linings (e.g., nasal, oral, vaginal, rectal, gastrointestinal mucosa, and the like).
  • suitable means include but are not limited to: inhalation (e.g. as a mist or spray), orally (e.g. as a pill, capsule, liquid, etc.), intravaginally, intranasally, rectally, etc.
  • the mode of administration is oral or by injection.
  • the compositions may be administered in conjunction with other treatment modalities such as substances that boost the immune system, various chemotherapeutic agents, antibiotic agents, and the like.
  • the dose of antibody that is administered varies according to factors such as the exact type of disease, the method of administration, overall health of the patient, etc., but is generally in the range of from about 1-100 mg/kg, or from about 2.5 -75 mg/kg or 5-50 mg/kg of body weight, including all whole number and decimal fractions thereof lying within the ranges.
  • the antibodies described herein are used to detect the expression level of IGFBP-3R (TMEM219) to diagnose and prognose cancer.
  • the antibodies may be labeled with a detectable reporter molecule.
  • a reporter molecule is defined herein as any moiety that may be detected using an assay.
  • Non-limiting examples of reporter molecules that may be conjugated to antibodies include but are not limited to enzymes, radiolabels, haptens, fluorescent labels, phosphorescent molecules, chemiluminescent molecules, chromophores, luminescent molecules, photoaffinity molecules, secondary or tertiary antibodies, and colored particles or ligands, such as biotin.
  • linking molecules may be used to join the antibody to the reporter molecule.
  • the antibodies may be immobilized on a solid support for use in an assay, e.g. on beads, in the wells of an assay plate, etc.
  • compositions for diagnostic agents can include any of the components listed above for compositions, but less care needs to be taken to promote physiological compatibility.
  • the assay solution is aqueous based and is buffered, and may contain preservatives, various salts, etc.
  • Kits comprising a container comprising the antibodies are also provided.
  • the kits may contain other reagents (e.g. reagents to detect a detectable label), directions for use, positive and/or negative reference standards, etc.
  • Diagnostic assays generally involve obtaining a biological sample of interest from a subject (e.g. a blood or plasma sample, a tissue sample, a biopsy sample, etc.) and exposing the sample to one or more antibodies as disclosed herein under conditions which allow binding to the antibody to a target molecule of interest, e.g. the TMEM219 molecule.
  • the antibodies generally comprise a detectable label, which after binding to the molecule of interest, is detected by methods known in the art and specific for each different label, and the amount of labeled antibody is correlated to the amount of the molecule of interest that is present in the sample, e.g. by the use of one or more reference values, as described below.
  • the methods described herein are used to diagnose cancer and/or to confirm a cancer diagnosis and/or to determine the prognosis of a patient with cancer, e.g. to predict one or more of metastatic potential, chances of recurrence and prospects for survival.
  • the patients treatment is tailored (modified, selected, etc.) according to the results of the method. Cancers which may be assessed in this manner include but are not limited to breast cancer, colon cancer, lung cancer, ovarian cancer, pancreatic cancer, liver cancer and leukemia.
  • a tumor sample e.g. a biopsy sample
  • a reference level of TMEM219 expression e.g. a reference level of TMEM219 expression
  • iii if the level of TMEM219 expression is the same or lower than the reference level of TMEM219 expression, then, iv) concluding that the patient has a poor prognosis and providing aggressive anti-cancer treatment to the patient, examples of which include combination therapy with TMEM219 agonist antibody and chemotherapy, radiotherapy, adjuvant therapy or hormone therapy or v) if the level of TMEM219 expression is higher than the reference level of TMEM219 expression, then iv) concluding that the patient has a good prognosis and providing less aggressive anti-cancer treatment to the patient, examples of which
  • the level of TMEM219 expression in the tumor sample is determined by any of many known methods for determining protein expression, including but not limited to: measuring the protein per se e.g. using antibodies (which are generally labelled with a detectable label), etc.; measuring mRNA encoding the protein, e.g. via PCR using primers (which are generally labelled with a detectable label), etc.
  • the level of TMEM219 expression is compared to at least one reference value.
  • suitable reference values is known in the art. Generally, such a value is established by measuring a substance of interest (protein, mRNA, etc.) that is indicative of the amount of TMEM219 expression in one or more appropriate control groups of comparable individuals who are healthy, i.e. in the present case, individuals who do not have cancer.
  • additional reference values may also be used, e.g. reference values established using tissue from other cancer patients with high and/or low levels of TMEM219 expression, and reference values based on cancerous and/or non-cancerous tissue from the patient him/herself.
  • Reference values may be obtained from patients with cancer or who have had cancer and who have or have not been treated for cancer, etc. e.g. patients in remission, those being actively treated for cancer, and the like. Controls may or may not be matched with respect to e.g. age, gender, ethnicity, overall health, life style, etc., as appropriate.
  • a level of TMEM219 expression that is "equal to”, “higher than” or “lower than” a reference value is: the same as the reference value (e.g. +/- about 5-10% of the reference value), or higher than the reference value e.g. at least about 10% or more higher than the reference value, or lower than the reference value e.g. at least about 10% or more lower than the reference value, respectively.
  • an amount of mRNA that is "equal to” is within +/- about 1-5% of Log [Fragments Per Kilobase of transcript per Million (RPKM)+1] 6.06 for colorectal cancer, 5.80 for ovarian cancer, 5.93 for pancreatic cancer, 5.43 for hepatocellular carcinoma, 5.77 for NSCLC and 5.91 for TNBC.
  • a "higher” level is a value that is at least about 5% greater that the above mentioned Log(RPKM+1) values.
  • a “lower” level is a value that is at least about 5% less than the above mentioned Log(RPKM+1) values.
  • an "equal" value falls within a range of from about 6.363 to about 5.757, a high value exceeds 6.363 and a low value is below 5.757.
  • the "equal" range may be within 1-10% of the indicated values, low values are at least 1-10% below and high values are at least 1-10% above the indicated reference values.
  • Patients that are found to have TMEM219 expression levels equal to or lower than a suitable corresponding reference value are considered to have a poor prognosis, e.g. a high likelihood of one or more of metastasis, recurrence, and low overall survival. Such patients are treated with an aggressive treatment regimen, as described below.
  • Patients that are found to have TMEM219 expression levels higher than the reference value are considered to have a good prognosis, e.g. a low likelihood of one or more of metastasis, recurrence, and overall high expectation of survival.
  • Such patients are treated with a less aggressive treatment regimen, as described below, and can avoid suffering the unwanted, detrimental side effects of aggressive treatment.
  • Such patients may in fact need no therapy (or no further therapy, if they have already been treated) but may benefit from monitoring the level of TMEM219 expression on an ongoing basis.
  • Therapeutic regimens may be comprised of the use of cancer chemotherapeutic agents and/or radiation and/or surgery.
  • a cancer chemotherapeutic agent is a chemical compound or biological agent that retards, slows, or stops the growth of cancer or is approved to treat cancer by the U.S. Food and Drug Administration.
  • cancer chemotherapeutic agents include, but are not limited to: paclitaxel, docetaxel, imatinib mesylate, sunitinib malate, cisplatin, etoposide, vinblastine, methotrexate, adriamycin, cyclophosphamide, doxorubicin, daunomycin, 5-fluoruracil, vincristine, endostatin, angiostatin, bevacizumab, and rituximab.
  • Another example of a cancer treatment agent is radiation.
  • the cancer treatment may comprise radiotherapy, fractionated radiotherapy, chemotherapy, or chemo-radiotherapy (a combination of one or more chemotherapeutic agents and radiation).
  • "Biological" anti-cancer agents include e.g. antibodies, proteins, RNA, siRNA, single guide RNA (sgRNA), DNA, etc.
  • an "aggressive cancer treatment” or “aggressive cancer treatment regimen” is generally determined by a medical professional such as a physician and/or radiologist and can be specific for each patient.
  • an aggressive cancer treatment regimen is as defined by the National Comprehensive Cancer Network (NCCN), and has been defined in the NCCN Guidelines TM as including one or more of 1) intensified imaging (CT scan, PET/CT, MRI, chest X-ray), 2) discussion and/or offering of sentinel lymph node biopsy with subsequent partial or complete lymphadenectomy, 3) inclusion in ongoing clinical trials, and 4) therapeutic intervention with interferon alpha treatment and radiation to nodal basin.
  • NCCN National Comprehensive Cancer Network
  • the phrase "aggressive cancer treatment” refers to a cancer treatment, or combination of treatments, and/or a chemotherapy regimen that is effective for treating the target cancer tumor or cell, but is associated with or known to cause higher toxicity and more side effects than another type of treatment for the specified cancer type.
  • Aggressive treatment may include one or more of surgical intervention, chemotherapy, radiation therapy, adjuvant therapy, hormone therapy, close clinical surveillance, etc.
  • Aggressive treatment may comprise proactive treatment to reduce or prevent metastasis, including distant or multiple metastases, e.g. using systemic chemotherapy.
  • exceptionally toxic chemotherapeutic agents may be preferred, as may higher and/or more frequent doses of one or more anti-cancer agents, and/or a longer course duration of therapy (chemotherapy, radiation, etc.), and/or a repetition of therapy.
  • chemotherapeutic agents may be preferred, as may higher and/or more frequent doses of one or more anti-cancer agents, and/or a longer course duration of therapy (chemotherapy, radiation, etc.), and/or a repetition of therapy.
  • chemotherapeutic agents may be preferred, as may higher and/or more frequent doses of one or more anti-cancer agents, and/or a longer course duration of therapy (chemotherapy, radiation, etc.), and/or a repetition of therapy.
  • a radical mastectomy may be recommended, together with lymph node removal, chemotherapy and radiation for a breast cancer patient.
  • Less aggressive treatment may also comprise surgical intervention, chemotherapy, radiation therapy, adjuvant therapy, hormone therapy, or close clinical surveillance, etc. It may also comprise proactive treatment to reduce or prevent local, organ-specific, tissue specific, or site-specific metastasis.
  • the treatment may be more localized and focus on the primary tumor, using e.g. resection followed by targeted drug therapy, such as treatment using antibodies which target the particular tumor type, or an implanted radiation source, etc.
  • a lumpectomy may be recommended to remove a cancerous breast tumor, preceded by neo-adjuvant treatment to shrink the tumor prior to surgery, rather than a radical mastectomy. If a course of radiation is prescribed, it may be a shorter and/or less intense course than that which is recommended for aggressive treatment.
  • a cancer treatment, combination of treatments, or chemotherapy regimen is less or more, and this may vary by cancer type, the age and general physical health of the patient, etc.
  • a less aggressive treatment may include adjuvant chemotherapy comprising surgical resection of the primary tumor and a chemotherapy regimen comprising 5-FU, leucovorin and bevacizumab
  • a more aggressive cancer treatment may include adjuvant chemotherapy comprising surgical resection and a chemotherapy regimen comprising FOLFOX and BV
  • the most aggressive cancer treatment may include surgical resection and a chemotherapy regime comprising Irinotecan and Cetuximab.
  • TMEM219 expression may benefit from monitoring TMEM219 expression on an ongoing basis.
  • subjects include but are not limited to subject with a genetic predisposition to develop cancer (e.g. women with mutations in one or both of the BRCA1 and BRCA2 genes), or who have experienced an environmental insult that may result in cancer (e.g. exposure to radiation, inhalation of toxic particles, contact with carcinogenic chemicals, etc.) or who have or are engaged in high risk activities with respect to cancer such as smoking.
  • Such subject may be monitored on an ongoing basis by determining TMEM219 expression levels over an extended period of time (months or years) and by a comparison of early, non-symptomatic levels in a tissue of interest (e.g. breast or lung tissue) to levels measured over time, or to a relevant reference value. In this manner, the development of cancerous tissues may be detected and early treatment can begin.
  • a tissue of interest e.g. breast or lung tissue
  • the present disclosure also provides IGFBP-3R agonists for use in the treatment of cancer.
  • the patient may or may not have been diagnosed using the methods described in the preceding section.
  • the agonists advantageously cause cancer cell death without killing normal, non-tumor cells.
  • the IGFBP-3R agonists are mAbs as disclosed herein, and the methods involve preventing or treating cancer by administering a therapeutically effective amount of at least one of agonist of the IGFBP-3R, such as a mAb disclosed herein.
  • the antibodies are used in single-agent therapy. In other aspects, the antibodies are used in combinatorial antitumor activity (which may give additive or synergistic results) with other chemotherapeutic agents including but are not limited to iniparib, gemcitabine, onartuzumab carboplatin, cisplatin, paclitaxel, bortezomib, erlotinib, everolimus, synribo, etoposide, doxorubicin, venetoclax, navitoclax, nivolumab and pembrolizumab.
  • chemotherapeutic agents including but are not limited to iniparib, gemcitabine, onartuzumab carboplatin, cisplatin, paclitaxel, bortezomib, erlotinib, everolimus, synribo, etoposide, doxorubicin, venetoclax, navitoclax, nivolumab
  • the agonists may be used in combination with other cancer therapies such as radiation, surgery/resection,
  • the cancer that is treated is breast cancer (e.g. TNBC), colon cancer, lung cancer, ovarian cancer, pancreatic cancer, head and neck cancer, prostate cancer, liver cancer or a liquid tumor (e.g. a leukemia), etc.
  • the amount of antibody that is administered is generally in the range of from about 1-100 mg/kg, and is preferably from about 5 to 50 mg/kg, e.g. about 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 mg/kg.
  • EXAMPLE 1 Diagnostics: TMEM219 as a molecular marker for predicting recurrence and survival in cancer
  • TMEM219 as a molecular marker for predicting recurrence and survival in breast, colon and lung cancer.
  • Oncotype DX a very expensive gene test, is routinely used in hospitals to predict chemotherapy benefit and recurrence risk of patients with breast, colon and prostate cancer.
  • the present TMEM219 assay is an excellent tool for replacing or complementing the Oncotype DX test.
  • Figure 4 depicts a schematic overview of the mechanism of action of the IGFBP-3/TMEM219 axis in cancer.
  • TMEM219 agonist does not in and of itself represent a useful therapeutic agent due to significant post-translational modification: proteolysis induced by tumor activated proteases attenuates TMEM219 antitumor signaling.
  • monoclonal antibodies mAbs that activate TMEM219, i.e. "agonist antibodies” were created.
  • the process of manufacturing mAbs has been standardized in the art and mAbs are known to exhibit robust stability within the body, without inducing major deleterious side effects.
  • the "agonist mAb” approach advantageously precludes the need to use toxic compounds to kill the cancer cells.
  • TMEM219 specific monoclonal antibodies was generated and two exemplary antibodies were sequenced (#245 and #274, Figure 5A and B). Further, a TMEM219 #274-human IgG1 chimera which acts like the TMEM219 natural agonist was also developed ( Figure 5C ). As shown in Figure 6A-C , treatment with one of the hybridoma cell-produced TMEM219 agonist mAbs [TMEM219 mAb#2 (#274)] significantly inhibited not only MCF-7 estrogen-responsive breast cancer cell growth but also MDA-MB231 TNBC cell growth.
  • MCF-10A normal immortalized mammary epithelial cell growth was not inhibited despite expressing a similar level of TMEM219 at the mRNA and protein levels. It was further observed that TMEM219 mAb#2 (#274) inhibits the growth of both breast cancer cell lines in a dose dependent manner with 70% growth inhibition at the concentration of 30nM (p ⁇ 0.001).
  • TMEM219 agonist mAbs have tumor suppressive activity not only on cancer cells in culture, but also in animal models of human cancer (models representing non-small cell lung cancer, triple-negative breast cancer (TNBC), colon cancer, and prostate cancer). In addition, TMEM219 agonist mAbs also suppress the tumor-activated signaling critical to tumor angiogenesis, metastasis and radio-/chemo-resistance. Of high importance is the fact that, despite strong anti-cancer cell activity, the TMEM219 agonist antibodies have no deleterious cell killing effect on normal, non-tumor cells.
  • TMEM219 agonist mAbs The antitumor and anti-metastatic effects of TMEM219 agonist mAbs was investigated using a bioluminescent orthotopic MDA-MB231 triple negative breast cancer (TNBC) mouse model.
  • TNBC triple negative breast cancer
  • TMEM219#274-hIgG1 chimera detects TMEM219 in MCF-7 cell lysates ( Fig. 7C , left panel) and also activates caspase-3 and inhibits tumor-activated NF-kappaB signaling in bioluminescent MDA-MB-231 cells ( Fig. 7C , right panel).
  • TMEM219 knockdown using CRISPR/Cas9 gene editing techniques resulted in complete abolishment of TMEM219 agonist mAb-induced cell growth inhibition ( FIG. 7D ).
  • sgRNA sequence targeted single guide RNA
  • TMEM219 agonist mAbs Treatment with 100nM TMEM219 agonist mAbs resulted in significant growth inhibition in the control (68% inhibition), whereas only 10% and 6% growth inhibition was observed in sgRNA-1 and sgRNA-2-transfected cells, respectively, after TMEM219 agonist mAbs treatment ( bottom panel ) .
  • TMEM219 agonist mAbs-induced anticancer effects are mediated through TMEM219 antitumor signaling in human cancers ( Fig. 7D ).
  • TMEM219 agonist mAbs were investigated using a bioluminescent orthotopic breast tumor mouse model.
  • MDA-MB231 cells expressing dtTomato- Luciferase were injected into the fourth mammary fat pad of 8 week old NOD-SCID-IL2yR-/- mice.
  • administration of a low dose of TMEM219 mAb (1mg/kg body weight) resulted in tumor shrinkage up to 25% (p ⁇ 0.01) at day 29 after tumor cell injection.
  • TMEM219 is readily detectable in all TNBC PDX cells tested at the protein and mRNA levels ( Fig. 9A and 9B ). Those expression levels were comparable to the established TNBC cells, MDA-MB231 and MDA-MB468.
  • immunohistochemistry data clearly demonstrate that TMEM219 is expressed in both PDX tumors and present mainly in cell membrane and cytoplasmic region but not in the nucleus ( Fig. 9C ).
  • TMEM219 agonist mAb#2 (#274) treatment resulted in a significant growth inhibition not only in chemodrug-sensitive WHIM30 but also in chemodrug-resistant WHIM2 PDX TNBC cells ( Fig. 9D ).
  • TMEM219 agonist mAb #274 (1mg/kg body weight) resulted in tumor shrinkage of up to 29% at day 30 after tumor cell injection ( Fig. 10A ).
  • Fig. 10B No apparent body weight or damage in major organs was observed in TMEM219 agonist mAb administrated mice ( Fig. 10B ).
  • the size and weight of tumors isolated from TMEM219 agonist mAb administrated mice was significantly reduced compared with those from mouse IgG administrated mice ( Fig. 10C ).
  • the 25% reduction of tumor weight observed in mAb administered mice was comparable to the 29% tumor shrinkage shown in Fig.
  • CAC Colon cancer
  • IHC immunohistochemistry
  • antitumor effect of AAT may be attributed to reduced TMEM219 natural agonist proteolysis, thereby enhancing TMEM219 natural agonist/TMEM219-mediated antitumor/anti-inflammatory, and further ameliorating neutrophil-activated cytokine function such as activation of the IL-1 ⁇ /IL-6 axis in CAC.
  • TMEM219 agonist mAbs Therapeutic potential of TMEM219 agonist mAbs was further tested for colorectal cancer using an AOM/DSS CAC mouse model.
  • the in vivo preclinical data clearly demonstrate that mice treated with TMEM219 agonist mAb#2 (#274) showed a dramatic suppression of tumor number and size ( Fig. 14A ).
  • TMEM219 natural agonist treatment treatment with TMEM219 agonist mAbs inhibits HT-29 colon cancer cell growth ( Fig. 14B ) and tumor-activated NF-kappa B signaling as shown decrease of phosphorylated-NF-kappa B and -I kappa B alpha ( Fig. 14C ).
  • TMEM219 natural agonist in NNKA, one of the derivatives, suppressed NF-kappaB activity and induced apoptosis whereas suppression of TMEM219 with its specific shRNA hindered TMEM219 natural agonist-induced suppression of NF- ⁇ B and induction of cell death.
  • TMEM219 natural agonist is mainly mediated via TMEM219 in NNKA cells.
  • IGFBP-3 itself does not constitute an excellent targeted therapy for lung cancer due to its significant degradation by tumor-induced proteases such as MMPs and ADAM28, thereby attenuating IGFBP-3's antitumor function ( Fig. 11 ).
  • these findings demonstrate that TMEM219 is a key antitumor signaling and a therapeutic target in NSCLC.
  • Insulin resistance represents a common metabolic derangement that contributes to the development of many obesity-related comorbidities including T2DM. Although it is generally established that low-grade adipose tissue inflammation contributes substantially to the burden of IR, the pathophysiology underlying the development of IR is complex and multifactorial. Thus, a clearer understanding of the mechanisms leading to obesity-associated IR is necessary to identify novel targets for the prevention and treatment of many IR-driven conditions such as T2DM.
  • visceral fat in obesity consisting primarily of adipocytes, secretes various pro-inflammatory adipokines such as tumor necrosis factor (TNF), leptin, visfatin, resistin, and interleukin (IL)-6 creating a state of local thus accelerating events leading to systemic IR, T2DM and metabolic syndrome.
  • TNF tumor necrosis factor
  • IL-6 interleukin-6
  • IGFBP-3 inhibits TNF-alpha-induced NF-kappa B activity through IGFBP-3R, thereby restoring insulin signaling and negating TNF-alpha-induced inhibition of glucose uptake in human primary adipocytes, suggesting that the IGFBP-3/IGFBP-3R system plays an important role in cytokine/adipokine-induced IR in visceral adipocytes. Furthermore, there is a decrease in functional intact IGFBP-3 levels and an increase in IGFBP-3 degradation (proteolysis) in the circulation of overweight and obese adolescents when compared with their non-obese counterpart.
  • IGFBP-3R agonist mAbs #245 and #274 and non-agonistic IGFBP-3R monoclonal antibodies (#C314) were employed in the presence of insulin and TNF-alpha in fully differentiated adipocytes.
  • IGFBP-3R agonist mAbs #245 and #274, but not non-agonistic mAb restored TNF-alpha-induced inhibition of glucose uptake in primary adipocytes.

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Claims (8)

  1. Un agoniste du récepteur de la protéine 3 de liaison au facteur de croissance analogue à l'insuline (IGFBP-3R), dans lequel ledit agoniste est un anticorps qui se lie à IGFBP-3R et l'active et
    comprend :
    - une région déterminante complémentaire de chaîne lourde 1 ayant la séquence GYTFTFNYG (SEQ NO : 25),
    - une région déterminante complémentaire de chaîne lourde 2 ayant la séquence INTYTRET (SÉQ NO : 27),
    - une région déterminante complémentaire de chaîne lourde 3 ayant la séquence ARGSTMYGLDK (SEQ NO : 29),
    - une région déterminante complémentaire de chaîne légère 1 ayant la séquence QNVGTN (SÉQ NO : 31),
    - une région déterminante complémentaire de chaîne légère 2 ayant la séquence SAS,
    - une région déterminante complémentaire de chaîne légère 3 ayant la séquence HQYNSYPLT (SÉQ NO : 33).
  2. Agoniste selon la revendication 1, dans lequel la chaîne lourde de la région variable dudit anticorps comprend la séquence identique à au moins 90 % à SEQ ID NO : 6 et dans laquelle la chaîne légère de la région variable dudit anticorps comprend la séquence identique à au moins 90 % à la SÉQ ID NO: 8.
  3. Agoniste selon la revendication 1 ou 2, dans lequel la chaîne lourde de la région variable dudit anticorps comprend la séquence telle que présentée dans la SEQ ID NO : 6 et dans laquelle la chaîne légère de la région variable dudit anticorps comprend la séquence telle que présentée dans la SEQ ID NO : 8.
  4. Un agoniste du récepteur de la protéine 3 de liaison au facteur de croissance analogue à l'insuline (IGFBP-3R), selon l'une quelconque des revendications 1 à 3, pour son utilisation dans une méthode de traitement du cancer exprimant IGFBP-3R.
  5. Agoniste pour son utilisation dans la méthode selon la revendication 4, dans lequel ledit cancer est cancer du sein, cancer du côlon, cancer du poumon, cancer des ovaires, cancer du pancréas, cancer du foie, cancer de la tête et du cou, cancer de la prostate ou tumeur liquide
  6. Agoniste pour son utilisation dans le procédé selon la revendication 5, dans lequel la tumeur liquide est une leucémie.
  7. Acides nucléiques isolés codant pour un agoniste selon l'une quelconque des revendications 1 à 3.
  8. Vecteur comprenant les séquences d'acide nucléique codantes selon la revendication 7.
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KR102594780B1 (ko) 2023-10-26
US20220152177A1 (en) 2022-05-19
US11179451B2 (en) 2021-11-23
US11786585B2 (en) 2023-10-17
KR20230154472A (ko) 2023-11-08
EP3534932A4 (fr) 2020-09-09
US20190270808A1 (en) 2019-09-05
CN110139661B (zh) 2024-06-11
JP2020500844A (ja) 2020-01-16
JP7036446B2 (ja) 2022-03-15
WO2018085252A1 (fr) 2018-05-11
KR20190075112A (ko) 2019-06-28
CN110139661A (zh) 2019-08-16

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